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Fraunhofer Institute for Molecular Biology and Applied Ecology

3 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study
Johannes Felix Buyel 1, Rainer Fischer 1,2
1Institute for Molecular Biotechnology, RWTH Aachen University, 2Institute for Molecular Biology and Applied Ecology, Fraunhofer Gesellschaft

We describe a design of experiments approach that can be used to determine and model the influence of transgene regulatory elements, plant growth and development parameters, and incubation conditions on the transient expression of monoclonal antibodies and reporter proteins in plants.

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Environment

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants
Megan Garvey 1, Johannes Klinger *1, Holger Klose *1, Rainer Fischer 2, Ulrich Commandeur 1
1Institute for Molecular Biotechnology, Bio7, RWTH Aachen University, 2Fraunhofer Institute for Molecular Biology and Applied Ecology

Tobacco plants were used to produce a fungal cellulase, TrCel5A, via a transient expression system. The expression could be monitored using a fluorescent fusion protein, and the protein activity was characterized post-expression.

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Biology

Comparison of Tobacco Host Cell Protein Removal Methods by Blanching Intact Plants or by Heat Treatment of Extracts
Johannes F. Buyel 1,2, Jürgen Hubbuch 3, Rainer Fischer 1,2
1Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e. V., 2Institute for Molecular Biotechnology, RWTH Aachen University, 3Department of Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT)

Three heat precipitation methods are presented that effectively remove more than 90% of host cell proteins (HCPs) from tobacco extracts prior to any other purification step. The plant HCPs irreversibly aggregate at temperatures above 60 °C.

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